Objective The purpose of this study was to compare the efficacy of autologous inactivatedbone segment and allogeneic bone in reconstruction of bone defect after resection of malignant bone tumors inlimbs. Methods The clinical data of 42 patients with malignant bone tumors of extremities who underwenttumor resection and inactivated autologous or allogenic bone reconstruction from March 2015 to December. 2017 were retrospectively analyzed. Musculoskeletal Tumor Society system（MSTS）and International Society of Limb Salvage（ISOLS）scoring system were used to evaluate and compare the functional and imaging results. Results The follow-up period ranged from 15 to 40 months，with an average of 29 months. At the end of follow-up，the MSTSaverage score of the treated group was 26（17 ～ 31），and the ISOLS average score was 27（23 ～ 32）；The MSTS average score of the treated group was 25（15 ～ 30），and the ISOLS average score was 28（24 ～ 30）. There was nosignificant difference in MSTS and ISOLS scores between the two groups Conclusion After resection of malignantbone tumors in extremities，autologous inactivated bone replantation and allogenic bone reconstruction have the same effect，and there is no significant difference in imaging evaluation.

Objective To investigate the clinical effects and complications of massive allograft boneconstruction in malignant tumor. Methods Retrospective analysis was conducted on eligible bone tumorpatients treated in the Affiliated Tumor Hospital of Zhengzhou University from 2007 to 2018. The study includedgeneral information of patients，primary disease，surgical site，length of reconstructed allograft bone，methods ofinternal fixation, postoperative graft healing，complications of internal fixation，and the ultimate survival statusof the graft. Results A total of 37 patients were included in the study，including 21 males，an average age of22.41（9 ～ 53）years. The incidence sites included 11 of proximal humerus，3 of radius and ulna，12 of femur，8 ofproximal tibia，and 3 of pelvis. Primary tumors included osteosarcoma in 25，giant cell tumor in 5，Ewing's sarcoma in 4，chondrosarcoma in 2，and other 1 patient. The operation methods were intercalary bone grafting in 17 and osteoarticular grafting in 20 patients. Internal fixation mainly included limited contact plates（LCP）or intramedullary nails. The operation time was（180 ～ 360）min，and the blood loss was（400 ～ 2600）ml. Postoperative wound complications included superficial infection in 2 patients，rejection reaction and sinus tract formation in 1 patient，and joint stiffness in 5 patients. Postoperative allograft bone healing without any additional intervention happened in 20 patients（54.05%），with the healing time of 6 ～ 24 months. Problems of bone healing appeared in 17 patients （45.95%）and 22 cases. The complications included nonunion（21.62%），graft resorption（including shaft andhumerus head，27.03%），and graft refracture（16.22%）. Postoperative limb function and joint range of motion can be competent for the patient's daily activities，6 cases（16.20%）of patients had tumor metastasis after surgery，and 7 cases（18.90%）had local recurrence. The 5-year survival rate was about 65% after surgery，and the tumor recurrence and metastasis were correlated with the bone graft pattern. Conclusion Homologous large segment allograft plays a certain role in bone tumor resected operation，which can achieve the purpose of bone reconstruction and bone union. But it has a high incidence of middle-long term complications，such as bone nonunion，boneresorption and postoperative re-fracture. It is recommended that the allograft bone should be protected by rigid with sufficient length of internal fixations. Removed of the internal fixations was not recommended after surgery.

Objective To observe the effect and prognosis of keratoplasty for the treatment ofkeratoconus. Methods Twenty-four cases（24 eyes）of keratoconus with keratoplasty from April 2016 to June 2017 were studied. Of all the patients，ten cases had received penetrating keratoplasty，two cases had received lamellar keratoplasty and 10 cases had received deep lamellar keratoplasty. The refractive state of the cornea，cornealgraft，the number of corneal endothelium and the incidence of complications were observed at 1 months，3 months，6 months and 1 years after the operation. Results Postoperative corrected visual acuity was significantly improved in all patients. 12 cases（12 eyes）raised 4 ～ 5 lines，6 cases（6 eyes）raised 6 ～ 8 lines，and 6 cases（6 eyes）raised above 9 lines. Rejection action was found in 1 eye after penetrating keratoplasty. Corneal grafts were transparentin other cases，and there was no postoperative complications such as corneal opacity and double anterior chamberafter operation. The number of corneal endothelial cells decreased to varying degrees after surgery，the maximum loss rate was 3 months，and then tended to be stable. Conclusion Keratoplasty is an effective method for the treatment of advanced keratoconus. Deep lamellar keratoplasty has a lower incidence of rejection reaction and no damage foranterior chamber. It provides a safe and effective choice for the treatment of keratoconus.

Objective To obtain α-1，3-galactosyltransferase/β-1，4 N-acetylgalactosaminyltransferase（GGTA1/β4GalNT2）double gene knockout porcine fetal fibroblasts（PFFs）by CRISPR/Cas9 system and lay the foundation forestablishing Bama miniature pigs with α-Gal and SD（a）antigen deficiency. MethodsSingle-guide RNA（sgRNA）targeting the third exon of pig GGTA1 and the eighth exon of pig β4GalNT2 weredesigned using online tools（http://crispr.mit.edu），respectively. The synthesized sgRNA were cloned into pX330 plasmid containing Cas9 skeleton. The cleavage efficiency of the Cas9/sgRNA plasmids was assessed by the T7EN1 enzyme digestion assay. The Cas9/sgRNA vectors wereco-transfected withaneomycin-expression plasmid（tdTomato）into the PFFs. G418 was used to screen the positive monoclonal cells and sanger sequencing was used to determine the genotypes of monoclonal cells. Results Cas9/sgRNA expression vectors targeting GGTA1 and β4GalNT2 gene were successfully constructed and transfected into PFFs. Among the 48 G418-resistant colonies obtained，five hadbiallelic modifications in both GGTA1 and β4GalNT2 loci. Conclusion The GGTA1/β4GalNT2 double gene knockout PFFs were successfully obtained by the highly efficient CRISPR/Cas9 targeting，which could contribute tothe generation of GGTA1/β4GalNT2-deficient Bama miniature pig models.

Objective To explore the application of knowledge-attitude-practice and life hope plan nursing intervention for the family members of renal transplant patients，to observe the influence of the family members' knowledge，beliefs and hopes and to assess the improvement of the patient's quality of life. MethodsFrom February 2016 to November 2017，58 patients and 116 families were enrolled，who were divided intotwo groups ：29 patients and 58 cases of family members in each group. The family members of control groupunderwent routine nursing intervention. Intervention group applied of knowledge-attitude-practice and life hope plan nursing intervention. Assessed the level of family members' knowledge and belief before and afterintervention，assessed the family members' hope level and self-efficacy by herth hope index（HHI）and the generalself-efficacy scale（GSES），and assessed the quality of life of patients. Results The family knowledge，beliefs，behavioral scores，total knowledge and belief scores，HHI scores，GSES score，and total quality of life of patients of the intervention group were（71.80±5.92）points，（54.61±5.26）points，（51.56±4.15）points，（174.59±8.96）points，（41.76±0.73）points，（34.80±0.64）points，（132.80±8.29）points，all higher were than the control group（62.25±5.81）points，（49.50±4.72）points，（46.89±4.03）points，（163.40±9.08）points，（36.29±0.80）points，（30.15±0.67）points，（124.46±8.03）points，the two groups had statistical differencesignificantly（P＜0.05）Conclusion The application of knowledge-attitude-practice and Life hope plan nursing intervention for the family members of renal transplant patients，the family members of the patients are well-informed，the hope level is significantly improved，the patient's quality of life is improved significantly，and the quality of careis significantly improved.